HU190910B - Method for producing glue suitable for fixing sorbent layer of pressurized one and multiple layer chromatography - Google Patents

Abstract

The invention relates to a method for producing a fine-grained (2-20 mm), required for the overprinting monolayer and multilayer chromatography, consisting of an organic and / or inorganic base material and / or provided with a bound or unbound apolar phase sorbent layers ( the particle size of which is mainly in a narrow range) suitable polyacrylamide adhesive, wherein from a mixture of aqueous solutions of acrylamide and N, N-methylene-bis-acrylamide under the action of catalysts, preferably, N, NN, N-tetramethyl -ethylene-diamine and ammonium peroxy-disulfate, in a temperature range between 21 and 30C, a polyacrylamide gel matrix is formed, from which the fraction suitable as an adhesive is precipitated after homogenization by a water-soluble solvent. The precipitate formed is diffused with water. The failing and spreading are repeated several times.

Description

(57) EXTRAS

The polyacrylamide adhesive produced by the process of the present invention is a silica gel sorbent with an organic and / or inorganic feedstock and / or a bonded or unbound apolar phase, by pressure monolayer and multilayer chromatography with a particle size of 2 to 20 µm and a particle size range of Δ um <2. suitable for fixing. The adhesive is prepared by acrylamide containing 1.6-2.5% by weight, preferably 1.9-2.2% by weight, and N, N'-methylenebis (acrylamide) 0.6- Catalysts known from a mixture of 1.0%, preferably 0.8-1.0% aqueous solutions - preferably Ν, Ν, Ν ', Ν'-tetrarrethylethylenediamine and diammonium peroxodisulphate - by precipitation from a polyacrylamide gel matrix formed at a temperature in the range of 21 ° C to 30 ° C after being homogenized with a water-miscible organic solvent, the unreacted monomers and oligomers are separated from the resulting polymer, the precipitate is washed with water and the pulp is repeated at least three times.

190 910

FIELD OF THE INVENTION This invention relates to a process for the production of a special-purpose polyacrylamide-based adhesive suitable for use in fine and mainly fine particle size organic materials (cellulose, polyamide, synthetic resin, etc.) and / or inorganic for high pressure monolayer and multilayer chromatography. suitable sorbents and their mixtures on different carrier sheets (silica gel, alumina, chemically bonded and / or non-polar silica silica, etc.).

Comparison of trends in laboratory instrumentation techniques today clearly shows a sharp rise in chromatographic techniques, which may be even more pronounced in the coming decades. This is especially true for columnar and planar liquid chromatography techniques. The so-called. High Performance Liquid Chromatography (HPLC) variants, due to their particular efficacy, are now leading in the analysis of a wide variety of groups of compounds, but also have significant preparative uses (e.g., H. Engelhardt, Hochdruck-Flüssigkeitschromatographie, 2. Auf. Verlag, Berlin-Heidelberg-New York, 1977). The development of HPLC and the results achieved with it have also brought about the need for a major overhaul of the most commonly used liquid chromatographic technique, liquid chromatography.

The disadvantage of classical layer chromatography [where a mobile phase (solvent mixture) migrates over the stationary phase (solvent mixture or sorbents) and carries one or more substances at different speeds) is that the development time is long and the separation is not always perfect due to the theoretical number of plates is limited.

Significant progress has been made in developing high performance layer chromatography (HPTLC) (A. Zlatkis and RE Kaiser, High Performance Thin-Layer Chromatography, Elsevier, 1977, and W. Bertsch, S. Hara, RE Kaiser and A. Zlatkis, eds.) (Instrumental HPTLC, Hüthig, Heidelberg, 1980), the most important features of which are the use of fine-grained sorbent sheets and instrumentation. However, the known fact that effective separation can be achieved with fine-grained (e.g. 5 µm) HPTLC also demonstrates the limitations of this technique, since good resolution can only be achieved at short distances, at longer development distances (e.g. 15-20 cm) - slowing solvent flow due to diffusion increases, spots (or bands) spread.

Thus, it was logical to think of a technique approaching well-standardized column chromatography, while retaining the indispensable advantages of layer chromatography (visual evaluation, aggressive reagents, map identification, etc.).

These needs are met by the so-called. pressurized layer chromatography (OPLC), which is classical layer chromatography (TLC), high performance layer chromatography (HPTLC) (this technique is based primarily on the use of fine particle layers) and high performance liquid column chromatography (HPLC). Its basic equipment is the pressurized ultramicro chamber, in which the sorbent layer (e.g. silica gel, alumina, talc, etc.) is advantageously suppressed by a pressurized water cushion system through a flexible membrane thereby eliminating the vapor space above the layer. The metering micropump feeds the eluent into the sorbent layer with a controlled pressure flow (for example, U.S. Patent No. 173,749; British Patent No. 1,570,760; and U.S. Patent No. 4,346,001). One-way, bidirectional, linear, circular, triangular (anticircular) development is possible by changing the location of the eluent inlets.

For effective linear OPLC, the edges of the sorbent layers need to be sealed, preferably by impregnation or plastic film, to prevent the eluent from leaking under overpressure. Migration of the eluent with a linear front can be achieved by providing a channel around the inlet of the eluent or by placing a wire (plastic sheet). This has recently been solved by placing a resilient pad (which otherwise protects the cushion, replaceable, etc.) on the sorbent layer on the motherboard, which has one or more channels for guiding the eluent around the inlet side of the eluent on the sorbent side. (Insertion sheet and / or insertion sheet system for overpressure for ultramicro chamber, Hungarian Patent Application No. 323/82.)

More recently, it has been found that a pressurized ultramicro chamber is capable of being developed on multiple sheets simultaneously if the sheets are specially trained for this purpose by perforating them in any shape and size around the point of application to the eluent layer. Perforated laminated sheets can be incorporated into a system in combination with a conventionally sealed edge laminated sheet, making this system suitable for different types of development and especially for the simultaneous examination of a large number of patterns. (Hungarian Patent Application No. 1335/82, Special Layer Base and / or Layer Base System for High Pressure Multilayer Chromatography.) This is high pressure multilayer chromatography.

A variety of base adhesives are used to fix the sorbent layer of ready-made layer bases used in classical layer chromatography (TLC) and high performance layer chromatography (HPTLC), particularly those that are resistant to aggressive reagents and relatively inert.

One of the groups of conventional adhesive materials used to date (e.g., agar-agar, starch, polyvinyl alcohol), polyvinyl acetate, is characterized by its sensitivity to aggressive reagents, particularly its partial solubility in aqueous systems, their application is now outdated (e.g. German Patent No. 1,442,446, pp. 7-8; Hungarian Patent Application No. 16T267).

Instead, they use inert organic polymer or inorganic adhesives, which are also resistant to aqueous systems, organic solvents and partially reagents. In German Patent No. 1442,446, acrylic and methacrylic acid or their salts or ethylene-maleic acid copolymers are used as binders. their salts are based on a high degree of polymerization. In German Patent No. 1517,929, various organic and inorganic based sorbent clog-23

190,910 using poly (acrylamide) and / or poly (methacrylamide) and their derivatives. German Patent No. 1,915,963 uses alkali metal silicate as a binder. However, these organic-based adhesives have the disadvantage that they are not always resistant to particularly aggressive reagents (e.g. sulfuric acid), especially at high development temperatures, and that an important reaction, such as the ninhydrin reaction, takes place with adhesives of a size such that quantitative evaluation is difficult. In the case of alkali metal silicate adhesive, it is difficult to ensure standard conditions.

These disadvantages, therefore, do not make these adhesives suitable for the production of all-purpose OPLC laminates, for which the following demands are made:

1. In pressurized monolayer and multilayer chromatography, the solvent mixture migrates by forced flow (via a dosing pump), so it is possible to use viscous solvent mixtures (aqueous buffers, etc.) and the adhesive or its fixed layer must be capable of viscous mixtures, cannot peel off, hump up, etc.

2. However, in a pressurized ultramicro chamber, the laminate is also subjected to significant external pressure, so the adhesive of the sorbent layer must withstand this pressure.

3. The moderate sensitivity of the adhesive to the ninhydrin reaction is particularly important, since high-pressure monolayer and multilayer chromatography is particularly advantageous in amino acid peptide quantitative analysis. The importance of detecting trace amounts of amino acids (such as N ^r -methylated lysines), which may sometimes be "lost" on the purple background, should be emphasized.

4. Requirements for high pressure monolayer and multilayer chromatography include solubilization of fine and narrow organic and inorganic based sorbents with fine particle size, and secure bonding of any mixture of organic and inorganic based sorbents, i.e., non-swelling, they must not interfere with bonding.

5. The great advantage of high-pressure monolayer and multilayer chromatography is that it is suitable for use with extremely fine-grained (e.g. 3-5 µm) and ultra-thin film-like (e.g. 45 µm thick) sorbent layer sheets, which provides better resolution than ever. it must also be capable of bonding sorbents.

6. The use of narrow particle size layer plates, irrespective of particle size, is a basic requirement in high pressure monolayer and multilayer chromatography, because of the uneven local flow leading to diffuse local flow between the particles of sorbents with significantly different particle size ranges. However, such sorbents are harder to fix than sorbent particles of different sizes, but the appropriate adhesive should also be suitable for this purpose.

7. One of the undoubted disadvantages of the finished laminate sheets over the column fillings is that the presence of the adhesive used to fix the sorbent layers reduces the development efficiency and, in particular, the reliability of the quantitative evaluation. Thus, an adhesive is required which, even in small amounts, provides the desired adhesion.

8. Finally, temperature has a particularly important role in the separation efficiency of high-pressure monolayer and multilayer chromatography, with optimum variations between systems, such as low and high temperatures.

Thus, it is desirable to provide an adhesive that is stable to both a suitable substrate [(e.g., aluminum foil) and a cover (e.g. Teflon)] at + 5 to + 120 ° C.

It is an object of the present invention to provide an adhesive which has overcome the drawbacks of the adhesives known and used to date for TLC and HPTLC sheets, and for the special needs described above for high pressure single and multiple chromatography.

The present invention is based on the discovery that the ability of various forms of polyacrylamide gels to coat the hair is the quality, quantity and matrix of the matrix forming monomers; monomers and oligomers remaining in the gel matrix adversely affect the polyacrylamide adhesive performance and indifferent surface properties, and must be removed from the poly (acrylamide) gel in order to achieve the intended purpose. It has also been found that the fraction suitable for bonding the sorbent layer can be precipitated from a homogeneous gel matrix in a controlled temperature range with a water-miscible solvent, preferably acetone, and can be easily separated from the monomers and oligomers which adversely affect adhesive properties.

The present invention relates to a process for the preparation of a polyacrylamide adhesive suitable for recording certain sorbent layers for high-pressure chromatography by solvent precipitation from a polyacrylamide gel matrix of a specific composition at a controlled temperature, followed by homogenization, followed by repeated precipitation and homogenization to obtain a non-articulating polyacrylamide gel adhesive.

The polyacrylamide gel adhesive produced by the process of the present invention is suitable for attaching pressure sorbent monolayer and multilayer chromatography layers. The adhesive has a density of between 1.004 and 1.008 g / ml, a viscosity of between 2 and 20, resistant to most solvents, waterproof, aggressive reagents (such as vanillin sulfuric acid), non-ninhydrin-sensitive, absorbent layers of high pressure they do not react under internal pressure or at higher development temperatures, retain their original form, and are also suitable for bonding fine and narrow organic and / or inorganic sorbents and mixtures thereof. Adhesive-bonded sorbent layers produced by the process of the present invention meet the requirements for quantitative evaluation better than sorbent layers made with prior art adhesives.

190 910

According to the invention, the acrylamide is 1.6-2.5%, preferably 1.9-2.2% and N, N'-methylenebis (acrylamide) 0.6%. To a solution of -1.0% by weight, preferably 0.8-1.0% by weight, of Ν, Ν, Ν ', N'-tetramethylethylenediamine (TEMED) followed by another catalyst, preferably diammonium peroxodisulfate was added at a temperature in the range of 21-30 ° C and the resulting polyacrylamide gel was homogenized in a homogenizer. This gives a liquid opalescent solution. This is followed by the addition of a water-miscible solvent, preferably acetone, to give a cottage cheese precipitate which, once the solvent is removed, is again pulped with the original amount of water and repeated several times as desired. The opalescent aqueous solution of the last curd precipitate in water is the adhesive solution.

The following describes preferred embodiments of the process of the invention.

A very preferred embodiment of the process according to the invention is starting with an aqueous solution of 2.0% by weight of acrylamide and 0.8% by weight of N, N'-methylenebis (acrylamide). Who.

The catalyst is preferably used in combination with Ν, Ν, Ν ', Ν'-tetramethylethylenediamine (TEMED) together with diammonium peroxodisulfate (NH ₄ ) ₂ S ₂ O ₈ , but other pairs of compounds such as (NH ₄ ) ₂ S may also be used as catalysts. ₂ O ₈ and 3- (dimethylamino) propionitrile or riboflavin and TEMED.

TEMED is added directly to the reaction mixture, while the diammonium peroxodisulfate is dissolved in a few ml of water and added to the middle of the vigorously premixed solution. The mixing stops spontaneously. Diammonium peroxodisulfate may also be added in solid form to the stirred solution. Here too, the mixing stops spontaneously. The ulceration begins after 4-5 minutes and usually ends after 30 minutes.

The gelation temperature is preferably selected to start at 22-23 ° C and rise to a maximum of 28-30 ° C.

The solidified gel is pulverized in a suitable pasteuriser such as Waring Blend for 5 sec and then the liquid opaque solution is suitable for solvent fractionation.

The opaque paste gel solution is mixed with water-miscible solvents such as acetone, methanol, ethanol and the like. we can precipitate the big molecules that are valuable to us and keep the small molecules in solution. The most preferred solvent, the acetone, is the most opaque solution to acetone: 1: 1 (v / v).

The acetone precipitated cottage cheese precipitate is preferably pulverized with water for 30 seconds in each case. Heat is added to the addition of acetone and the solution is decanted by cooling.

The quality of the adhesive is better the more times we precipitate it, but it is advisable to conduct 3 precipitations.

The polyacrylamide content of the opal adhesive solution ranges from 1.6 to 2.2% by weight. With this in mind, calculate the percentage of adhesive per sorbent, preferably between 0.5 and 5.0% by weight, depending on the sorbent, layer thickness.

The following examples illustrate the invention without limiting it.

Example 1

A. Preparation of Poly (Acrylamide) Adhesive G acrylamide and 8 g N, N'-methylene bis (acrylamide) were dissolved in 1000 ml distilled water. TEMED (3 ml) was added and the aqueous solution adjusted to 22.5 ° C was stirred vigorously in one direction, and 0.8 g (NH ₄ ) ₂ S ₂ O ₈ 0 in water (3 ml) was added to the center of the stirred solution. After 4 minutes, opalescence begins, with the temperature rising to 29 ° C, and after 30 minutes the gelation is complete.

The gel was pulverized in Waring Blend for 5 sec and 1000 ml of acetone was added to 1000 ml of opaque solution. Decant the solution by cooling and collect the collected cottage cheese precipitate, together with the precipitate deposited on the vessel wall, with 1000 ml of distilled water and pulp for 30 seconds. To the resulting opaque solution was added another 1000 ml of acetone, and the above operations were repeated, i.e., an additional acetone was precipitated. (The solutions obtained by decantation are always discarded.) Precipitation with ethyl alcohol or isopropyl alcohol gives similar results to acetone precipitation.

The 1000 ml opaque solution obtained in the last step had a polyacrylamide content of 0.021 g / ml, a density of 1.004 g / ml and a relative viscosity of 4.65.

B) Preparation of the sorbent layers

Using the adhesive prepared in A), the following layer was applied: To 57 g of glue (2.7% by weight) was added to 44 g of silica gel having an average particle size of 6 µm and an internal diameter of 910 ⁹ m. The resulting pulp was gently mixed and pulverized in Waring Blend for 60 sec. The resulting sour cream paste was applied to 0.1 mm thick aluminum foil by continuous lubrication at 0.2 mm thickness to give a 0.16 mm thick layer after drying. After edging, 7 x 20 x 20 cm sheets were obtained.

C) Development of high pressure single layer chromatography

The sorbent was scraped off on three sides of the sheets made according to point B at a width of 5 mm and a water and solvent-resistant plastic film was formed using a plastic dispersion (containing polyvinyl acetate) and poly (acrylamide). 15 cm of each color test of a mixture of indophenol, butter yellow and methyl red were placed 3.0 cm from the edge, 9-9 mm apart, and after drying of the sample application sites, the sheet was placed in a Teflon pressurized ultramicro chamber equipped with a solvent channel guide pad. 1.2 MPa, solvent methylene chloride, solvent flow rate 185 mL / h, development time 4.6 min The resulting chromatogram was evaluated by spectrophotometer and found to be ± 1.5% per color.

D) Development of high pressure multilayer (four layer) chromatography

Four of the laminated sheets prepared in (B) were impregnated with a water dispersion and solvent resistant plastic film at a width of 5 to 5 mm at the edges using a plastic dispersion and heated for 10 minutes at 100 ° C. The top 3 of the impregnated edges of the plywood sheet were perforated to a odd sealed edge 15 mm wide by 185 mm wide with a 2 mm cross section. The 3 pieces

190.910 aliquots of petroleum ether extract (3x15 pieces) made from inflorescence drug of chamomile compound 15 mm from the perforated gap were made on a perforated layer plate made in exactly the same way. After the sample application sites had dried, the sheets were superimposed on each other, and a fourth sheet with non-perforated but impregnated edges was placed at the bottom, which was also provided with samples from 15 other chamomile fragments. The system thus assembled was placed in a CHROMPRES 10 pressurized ultramicro chamber. Benzene was used for the development, the cushion pressure was 1.1 mPa and the development time was 16.2 minutes. After development, the plates were disassembled, dried, and then sprayed with 90% sulfuric acid (0.2% vanillin), heated at 105 ° C for 5 minutes, and evaluated visually using a chromatogram spectrophotometer. The sheets had the same front spacing and the same R 1 values, with standard deviation within the allowable range.

Example 2

Similarly to Example 1, the layers of silica gel lubricated were cut into 20 x 40 cm pieces. From the opposite sides of these layers, the sorbent was scraped to a width of 5 to 5 mm, whereby a plastic dispersion was used to form a water- and solvent-resistant plastic film for bidirectional high pressure chromatography. A 180 mm wide channel for solvent guidance was formed in the center of the layer sheet, while 15 aliquots of 15-15 different wheat kernel hydrolyzates were applied to both sides of the channel for 15-15 mm and 9-9 mm apart. After drying the sample application sites, the layer was placed in a CHROMPRES 10 pressurized ultramicro chamber. The cushion pressure was 1.2 MPa, the developing mixture (n-butanol) glacial acetic acid water 4: 1: 1 vol. ratio. Development time was 68 minutes.

After development, the dried slides were blown in with 0.2% methanol-acetic acid, copper sulfate-saturated ninhydrin reagent, heated, and the color chromatogram evaluated with a spectrophotometer.

Example 3

Using the adhesive described in Example 1, the following layer was applied: 40 g of talc with an average particle size of 3-4 μιη was diluted in 40 ml of distilled water with 40 ml of glue for 75 seconds in Waring Blend and the resulting sour cream pulp was 0.15 mm thick terephthalate plastic by continuous lubrication at 0.15 mm thickness, resulting in a 0.1 mm thick talcum layer after drying. After edging, 6 x 20 x 20 cm plywood sheets were obtained. The sorbent was scraped off on three sides of the layer sheets so prepared, and a water dispersion and solvent resistant plastic film was formed using a plastic dispersion (containing polyvinyl acetate) and poly (acrylamide). One of the impregnated edges of the finished talc layer sheets was applied with aliquots of Digital Plant Extracts and Authentic Digital Heart Glycosides from methanolic chloroform solution. The slab was placed in a CHROMPRES 10 ultramicro chamber. A developing solvent (methylethylketone) water (18:85 v / v) was used. The development time was 25 minutes.

After drying, the layer was blown with 20% aqueous phosphoric acid and heated at 120 ° C for 20 minutes to give yellowish-green spots, which were evaluated qualitatively and quantitatively.

EXAMPLE 4 By using the adhesive described in Example 1, a concentrated zonal layer was formed: 33 g of silica gel having an average particle size of 6 µm and an internal diameter of 9 · 10 ⁹ m, and diluted with 66 ml of distilled water in a larger container of a specially designed lubricating head A slurry of 43 ml of adhesive (2.71% silica) was poured by appropriate paste. While in the smaller container (with a smaller lubricating surface) 11 g of silica having an average particle size of 15 μτα and

15 ml of glue paste diluted with 21.5 ml of water were measured. The two pulps were applied to 0.1 mm thick aluminum foil by continuous lubrication at 0.1 mm thickness to give, after drying, a 0.08 mm thick concentric zone layer of 4 cm wide inactive diatomaceous earth and 16 cm wide active fine grain silica.

After edging, 10 x 20 cm x 20 cm layer sheets were obtained, which were extruded for high pressure monolayer chromatography as described in Example 1. Chamomile oil samples of different origins were used for separation, the components of which (polynols, s'.quiterpene alcohols, etc.) differ in sharp bands.

Example 5

Using the adhesive described in Example 1, a silicone film and resin mixed layer was prepared by pulping 28 g of silica gel on average particle size and 10 g of a strong cation exchange resin having a mean particle size of 58 µm with 42.5 ml + 70 ml of distilled water for 60 sec. The pulp was applied to 0.1 mm thick aluminum foil by continuous lubrication at 0.2 mm thickness to give a smooth, light-yellow layer of 0.17 mm, which resulted in 7 sheets of '0x20 cm'.

For pressurized monolayer and multilayer chromatography, the edges of the layer sheets were impregnated as described in Example 1, respectively. perforated.

To demonstrate the effectiveness of the sheets, hydrochloric acid hydrolysates of various plant proteins and authentic amino acid mixtures were applied. After drying, the layer sheet was placed in a CHROMPRES 10 chamber. Citrate buffer, pH 3.45, was used as the developing mixture, while 0.2% methanol-acetic acid ninhydrin reagent was used as the developing reagent.

Example 6 g acrylamide and 8 g N, N'-methylene bis (acrylamide) were dissolved in 1000 ml distilled water. TEMED (2.8 mL) was added and the aqueous solution adjusted to 23 ° C was thoroughly mixed in one direction and 1.9 g of crystalline (NH ₄ ) ₂ S ₂ O _g was sprayed into the center of the stirred solution. The ulceration begins after 5 minutes and the complete gelling is completed after 30 minutes.

The gel is further processed as described in Example 1 below. The resulting opal adhesive solution had a density of 1.005 g / ml and a viscosity of 6.8.

Using this adhesive, apply the following layer:

g of silica gel with an average particle size of 3 μπι and an internal diameter of 6 · 10 ^-9 m diluted in 56 ml with 90 ml water

-5190.9'Ο Pasteurized in Waring Blendor. The pulp was lubricated with 0.1 mm thick aluminum foil by continuous lubrication at 0.1 mm thickness to give 0.06 mm thick layer after drying. The edged, cut layers were wind-impregnated as described in Example 1 and used for coloring in a pressurized single-layer chromatography system, particularly with small amounts of sample (eg 5-10 mg) applied to the layer.

Example 7

Using the adhesive described in Example 6, the following layer was prepared:

Aluminum oxide with 10 µm average particle size was pulverized with a mixture of 39 ml of water and 28 ml of glue for 225 sec, and the pulp was spread on 2 mm thick 20 x 20 cm glass sheets with a 0.2 mm gap to give a 0.16 mm thickness after drying. a layer of alumina was obtained.

The edges of the laminated sheet activated at 110 ° C were scraped on three sides to a width of 4-4 mm and a plastic film was formed therein to form a sealing film. Aliquots of a 1: 1 solution of methanolic chloroform in purified extracts of poppy seeds were applied to the thus formed sheet 3 cm from the odd impregnated edge. After drying the sample application sites, the sheet was placed in a CHROMPRES 10 ultramicro chamber with a suitable plastic protective frame (to protect the pressurized ultra-microfiche water cushion system against the cleavage effect of the glass edges). The cushion pressure was 1.2 MPa. Eluent was eluted with a mixture of heptane-chloroform-ether (40:50:10 by volume) and, after development, a Dragendorff reagent modified by Vágújfalvi was used.

Example 8

Using the adhesive described in Example 6, layers similar to the silica gel layer applied therein were applied from a pulp supplemented with 1.8% Mn activated zinc silicate UV indicator per silica gel.

Example 9 g acrylamide and 16.2 g N, N'-methylene bis (acrylamide) were dissolved in 2000 ml distilled water. 4 ml of 3- (dimethylamino) propionitrile is added and the aqueous solution adjusted to 22 45 ° C is thoroughly mixed in one direction, and 1.0 g of (NH ₄ ) ₂ S ₂ O _g is rapidly dissolved in 3 ml of distilled water. and pour the solution into the center of the stirred solution. The ulceration begins after 8 minutes and the complete gelling is completed after 45 minutes. 50

The resulting gel was worked up as described in Example 1. The resulting opal adhesive had a density of 1.006 g / ml and a relative viscosity of 4.3.

Using this adhesive, the following layer was lubricated: 30 g of high purity cellulose having a mean particle size of 5 µm, was applied to a 0.9 mm thick aluminum foil with a thickness of 0.15 mm and a thickness of 0.15 mm in 30 ml of distilled water.

The edged, cut sheets were edge impregnated as described in Example 1 and used for amino acid analysis in a pressurized single-layer chromatography system. The developing mixture was (methyl ethyl ketone) acetonitrile acetic acid / water (3: 4: 1: 2 v / v). The cushion had an external pressure of 1.4 MPa and a development time of 59 minutes (in the case of over-run).

Example 10

The adhesive described in Example 9 was used to attach chemically bonded stationary phase silica gel (RP-18 silica gel) to aluminum foil:

g of C-18 reverse-phase silica gel powder was mixed with 80 ml of polyacrylamide adhesive and 60 ml of isopropyl alcohol and pulp for 175 sec. The resulting diluted pulp was smeared with 0.12 mm thick aluminum foil at a thickness of 0.18 mm. The dried laminates were used in a 1C (after cutting, edge impregnation) CHROMPRES 10 ultramicro chamber for reverse phase separation of poppy alkaloids in a 6: 4 v / v mixture of (acetonitrile) (0.005 m KH ₂ PO ₄ solution). The development reagent used was Dragendorff's reagent in ethyl acetate.

Example 11

Using the adhesive described in Example 9, the following preparative layers are applied:

88 g of 5 .mu.m average particle size, 6 - diluted 10 ¹¹ m internal pore silica with 175 ml of water 135 ml of adhesive 85 sec in a Waring blender slurried. The resulting pulp was 0.11 mm thick with degreased aluminum foil with 0.65 mm thick needles to give, after drying, a 0.5 mm layer of silica gel, which was N'-methylated by a spontaneous methylation and formylation reaction between E-lysine and formaldehyde. was used to prepare lysines. Distilled water is used as the eluent. The cushion pressure was 1.3 MPa.

Claims

Claims (4)

tographic silica gel sorbent layers of organic and / or inorganic base material and / or bound or unbound apolar phase with a particle size range of 2 to 20 µm 40 suitable for the preparation of polyacrylamide adhesive, characterized in that the acrylamide is 1.6-2.5% by weight, preferably 1.9-2.2% by weight of N, N'-methylene bis Known catalysts, preferably from Ν, Ν, Ν ', Ν'-tetramethylethylene, in a mixture of aqueous solutions of (acrylamide) 0.6-1.0% by weight, preferably 0.8-1.0% by weight diamine and diammonium peroxodisulfate - after homogenization in a polyacrylamide gel matrix formed at a temperature in the range of 21 ° C to 30 ° C, the unreacted monomers and oligomers are separated from the resulting polymer by precipitation with water. pulping and repeating the precipitation and pulping at least three times. 2. The process of claim 1 wherein the catalyst is diammonium peroxodisulfate and 3- (dimethylamino) propionitrile. 3. A process according to claim 1, wherein the catalysts are gauged at 22 ° C. 4. A process according to claim 1 wherein the unreacted monomers and oligomers are separated from the gel matrix by the resulting polymer.